Detection of tumor-reactive antibodies in the B16 melanoma model
| Mice . | % reactivity with B16-EpCAM cells (sera on day 0) . | % reactivity with B16-EpCAM cells (sera immediately before death) . | % reactivity with B16-EpCAM cells (sera on day 143)* . | mIgG2a† . | mIgG1† . |
|---|---|---|---|---|---|
| 1 | 5 | 87 | + | − | |
| 2 | 6 | 42 | + | − | |
| 3 | 5 | 70 | + | − | |
| 4 | 4 | 73 | + | − | |
| 5 | 5 | 30 | + | − | |
| 6 | 5 | 66 | + | − | |
| 7 | 5 | 57 | + | − | |
| 8 | 6 | 71 | + | − | |
| 9 | 5 | 47 | + | − | |
| 10 | 5 | 49 | + | − | |
| 11 | 6 | 69 | + | − | |
| 12 | 5 | 40 | + | − | |
| 13 | 5 | 54 | + | − | |
| 14 | ND | ND | ND | ND | |
| 15 | 4 | 8 | − | − | |
| 16 | 5 | 14 | − | − | |
| 17 | 6 | 10 | − | − | |
| 18 | 5 | 18 | + | − | |
| Control serum | 3 | 4 | 2 | − | − |
| Mice . | % reactivity with B16-EpCAM cells (sera on day 0) . | % reactivity with B16-EpCAM cells (sera immediately before death) . | % reactivity with B16-EpCAM cells (sera on day 143)* . | mIgG2a† . | mIgG1† . |
|---|---|---|---|---|---|
| 1 | 5 | 87 | + | − | |
| 2 | 6 | 42 | + | − | |
| 3 | 5 | 70 | + | − | |
| 4 | 4 | 73 | + | − | |
| 5 | 5 | 30 | + | − | |
| 6 | 5 | 66 | + | − | |
| 7 | 5 | 57 | + | − | |
| 8 | 6 | 71 | + | − | |
| 9 | 5 | 47 | + | − | |
| 10 | 5 | 49 | + | − | |
| 11 | 6 | 69 | + | − | |
| 12 | 5 | 40 | + | − | |
| 13 | 5 | 54 | + | − | |
| 14 | ND | ND | ND | ND | |
| 15 | 4 | 8 | − | − | |
| 16 | 5 | 14 | − | − | |
| 17 | 6 | 10 | − | − | |
| 18 | 5 | 18 | + | − | |
| Control serum | 3 | 4 | 2 | − | − |
EpCAM indicates epithelial cell adhesion molecule; IgG, immunoglobulin G; ND, not done.
Measurements were performed by flow cytometry. Tumor-reactive antibodies were detected with polyclonal fluorescein isothiocyanate-conjugated rat antimouse IgG antibody. Numbers represent percentage of positively stained cells.
Measurements of tumor-reactive sera were done as described in above footnote, but detection followed with IgG subclass-specific antibodies against mouse IgG2a and mouse IgG1.